CN1067249A - Amidinophenylalanine derivatives and their preparation and use - Google Patents

Abstract

The present invention describes amidinophenylalanine derivatives of formula I, Synthetic methods of these compounds, their use and pharmaceutical preparations containing these compounds.

Description

The present invention relates to amidinophenylalanine derivatives, the synthesis of these compounds, their application and pharmaceutical preparations containing these compounds.

A range of pathophysiological states are known to lead to depletion of antithrombin III (ATIII), the most important thrombin inhibitor in plasma. A decrease in ATⅢ increases the risk of thrombosis, as is known to occur with congenital ATⅢ deficiency. AT Ⅲ dropped below 75% of the normal value, will cause thrombotic complications. This complication often occurs in the form of disseminated intravascular masses after surgery or in a state of shock. As a result, life-threatening blood clots occur in many cases. Anticoagulants with different modes of action have hitherto been used in medicine for the treatment and prophylaxis of thrombotic disorders. To rescue the thrombotic danger, substances such as ATIII, heparin and more recently hirudin are used. Coumarin and indanedione derivatives for long-term prophylaxis. However, the anticoagulants mentioned all have in some cases significant disadvantages. Heparin, for example, can only be administered parenterally due to its polysaccharide structure, and its action also depends on the level of effective antithrombin III. Coumarins directly antagonize protein biosynthesis, and through protein biosynthesis, coagulation factors II, VII, IX, and X that depend on vitamin K cannot be fully provided, thus reducing the possibility of clot formation. So exhibit a brief delay in action. Known side effects are hemorrhagic skin necrosis, nausea and hair loss.

In contrast, low molecular weight thrombin inhibitors have the advantage of acting independently on thrombin directly as a cofactor, which binds directly to the active center of the enzyme and thus almost completely inhibits the enzyme. These substances are orally available due to their chemical structure.

Amino acid derivatives based on arginine and amidinophenylalanine are particularly well known. Compounds belonging to the first group such as D-phenylalanyl-L-prolyl-argininaldehyde and (2R,4R)-4-methyl-1-[N2-(3-methyl-1,2 , 3,4-Tetrahydro-8-quinolinesulfonyl)-L-arginyl]-2-piperidinecarboxylic acid monohydrate ("MD805"). MD805 is a specific competitive inhibitor of thrombin and has been used as a therapeutic drug. Another known amidinophenylalanine derivative is β-naphthalenesulfonyl-glycyl-R,S-4-amidinophenylalanylpiperidine (NAPAP). Derivatives of NAPAP are described in EP0236163 and EP0236164. That is, glycine (NH ₂ -CH ₂ -COOH) is replaced by another amino acid with the structure (NH ₂ -CHR1-COOH), where R1 is lower alkyl, lower hydroxyalkyl, phenyl or 4-hydroxy phenyl. 4-Amidinophenylalanine (Aph) can be methylated to N-methyl-Aph. In addition, various derivatives on the arylsulfo group of NAPAP, on the "bridge"-glycine and on the piperidine ring are described, most suitably having an α- or β-naphthalenesulfonyl group at the N-terminus, while Heterocyclic arylsulfonyl groups such as 8-quinolinesulfonyl are an order of magnitude less effective. Another disadvantage of these compounds, especially of the NAPAP structure type, is their lesser tolerability, unfavorable pharmacokinetic properties and partly lower specificity, especially the strong anti- Trypsin action. Substances containing p-amidinophenylalanine have the disadvantageous fact that they are not stable to enzymes in the intestine and liver when administered orally, so that these substances are rapidly metabolized in the intestine and liver. Replacing the piperidine ring in NAPAP with proline can improve tolerance and appropriate pharmacokinetics. However, a very significant disadvantage of the proline compound concerns its loss of thrombin inhibition about 100-fold compared to NAPAP. So far, the specificity of compounds for thrombin and trypsin has not been significantly improved by introducing substituents.

The object of the present invention is to provide novel compounds based on amidinophenylalanine, whose antithrombotic effect exceeds that of known compounds, which are highly resistant to enzymolysis, have improved tolerance, improved specificity and good pharmacokinetics.

The content of the present invention is the compound of formula I,

In the formula

R' is a naphthalene ring attached to the α-position or the β-position, optionally substituted by an alkyl group containing up to 3 carbon atoms and/or an alkoxy group containing up to 3 carbon atoms; or is a tetrahydronaphthalene ring or an indene ring attached to the α- or β-position, and the ring may optionally contain an alkyl group with up to 3 carbon atoms, and/or an alkoxy group with up to 3 carbon atoms; or It is a benzene ring, optionally containing an alkyl group with up to 4 carbon atoms on the benzene ring, and/or having up to 3 groups with the structure O-X, where O is oxygen, X is hydrogen, methyl, ethyl, n-propyl, isopropyl or tert-butyl, and/or a group having the structure -COOY, Y is hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, isobutyl, Isopentyl or neopentyl; or chroman ring system, the ring can have up to 5 alkyl groups, and the alkyl group can have up to 3 carbon atoms;

R1 is a group with the structure AB, A=-(CH ₂ )n-, n=1-4, B is an acid functional group, which can be a carboxylic acid group, which can be optionally esterified or become an amide, ester alcohol It can contain up to 17 carbon atoms; it can be a sulfonic acid group, a phosphoric acid group, a boronic acid group or a tetrazolyl group; R1 or a group with the structure ABC, A has the same meaning as above, B is a carbonyl or sulfonyl group, and C is formed by N - linked α, β, γ or δ amino acids or groups linked to aldonic acid via N-glycosidic bonds;

R2 and R3 may be the same or different, and they are alkyl groups with up to 4 carbon atoms, or R2 and R3 together form a heterocyclic ring with a maximum of 8-membered rings, and the rings may be substituted by hydroxyl groups or the number of carbon atoms may be up to 3 The hydroxyalkyl group is substituted, the hydroxyl group can be optionally esterified, and the corresponding carboxylic acid of the esterification can contain up to 17 carbon atoms, * represents a carbon atom in the R or S configuration, and the R-configuration is preferred.

These novel compounds are characterized in that the amino acid R1 in formula I is an acidic amino acid. Acidic amino acids are, for example, those found in proteins, glutamic acid and aspartic acid or also sulfoalanine, but also unnatural amino acids such as aminoadipic acid or 3-phosphorylalanine or 2- Amino-3-boronylalanine. Because these amino acids have an asymmetrically substituted carbon atom to become a chiral substance, new compounds made with these amino acids will also have different effects, and the final products made of corresponding L-configuration amino acids are highly active. many.

The characteristic of the group A in the structure R1 is preferably that n is 1 or 2, especially 1 is preferred. If the group C is an amino acid, it is preferably an α-, β-, γ-, or δ-amino acid. Examples are α-aminoadipic acid, α-aminobutyric acid, γ-aminobutyric acid, 4-aminobenzoic acid, 2-aminobenzoic acid, ε-aminocaproic acid, 1-aminocyclohexanoic acid, 2-aminocyclo Hexanoic acid, 3-aminocyclohexanoic acid, 4-aminocyclohexanoic acid, 1-aminocyclopentanoic acid, 2-amino-4,5-dimethylphenoxazinone(3)-1,8-dicarboxylic acid , 2-amino-3-hydroxy-4-methylbenzoic acid, α-aminoisobutyric acid, aminohydroxybutyric acid, β-aminoisobutyric acid, alanine, β-alanine, dehydroalanine , C-allylglycine, alliin, 2-amino-3-methyl-butyl-1,3-thiazolidine-5-carboxylic acid, 6-aminopenicillanic acid, α-amino-heptene Acid, 1-aminocyclopropionic acid, asparagine, aspartic acid, α-aminosuberic acid, azetidinic acid, aziridine, betonine, C-benzylphenylalanine , canavanine, citrulline, cysteine, cystathionine, lysoline, 3,4-dihydroxyphenylalanine, 4-sulfonylphenylalanine, 2,2-dimethyl Glyco-1,3-thiazolidine-4-carboxylic acid, felinine, glutamine, glutamic acid, glycine, hexahydronicotinic acid, homocysteine, histidine, homoserine, delta-hydroxy Lysine, γ-Hydroxyproline, β-Hydroxyproline, Isoleucine, Isoserine, Isovaline, Kynurenine, Lanthionine, Mimosine, Leucine, Lysergotine, Methionine, Mimosaline, Pyrrolic Acid, Norleucine, Norvaline, Injectionine, 2-Pipicolic Acid, Penicillamine, Phenylalanine, C-Phenylglycine, Picolinic Acid , proline, dehydroproline, β-phenylserine, pyridine-2-alanine, 5-pyrrolidone-2-carboxylic acid, 3-pyrazolealanine, quinoxaline-2-carboxylic acid , Rosine, Sarcosine, Selenocysteine, Selenomethionine, Serine, Statin, (1,3-Thiazolyl(2))-Alanine, β-(2-Thiazolyl)-Propanine amino acid, threonine, thyronine, thyroxine, 1,3-thiazolidine-2-carboxylic acid, tert-leucine, tryptophan, tryptathionin, tyrosine, valine. These amino acids can occur in the optically active D- or L-configuration.

If C is an aminodicarboxylic acid, such as D- or L-glutamic acid or aspartic acid, one of the carboxyl groups can be derivatized, especially the α-carboxyl group is preferably derivatized. These derivatives are preferably amides, for example amides of ammonia, methylamine, dimethylamine, benzylamine, piperidine or morpholine.

If the group C is an N-glycosidically linked aldonic acid, such compounds have the general formula:

X1 = -H, -OH, -CH ₃ , O-acetyl

X2 = -H, -OH, -CH ₃ , O-acetyl

X3 = -H, -OH, -CH ₃ , O-acetyl

These new compounds are unexpectedly very potent inhibitors of thrombin. Stability to intestinal and liver homogenates was also surprisingly found, and also a decisive improvement to trypsin and chymotrypsin compared to known compounds.

It has also surprisingly been found that the R' group of the compounds according to the invention has a decisive influence on the potency and specificity of action. When R'=β-naphthalenesulfonyl and R1=aspartic acid of the compound, the inhibitory effect on thrombin and trypsin is very similar, so the specificity is low. When introducing alkyl and/or hydroxyalkyl and/or hydroxyl and/or carboxyl groups on the sulfonic acid group, groups with up to 3 carbon atoms are preferred, especially methyl and/or methoxy Base time, can significantly improve the selectivity to thrombin. This means that after substitution with one or more of these groups, the properties of the resulting substances show increased inhibition of thrombin and at the same time reduced inhibition of trypsin.

Thus, the properties of this class of important substances as thrombin inhibitors can be decisively influenced by the derivatization modes described here, leading to superior compounds.

Particularly effective compounds of the present invention (determinable from the Ki value) are those of the structure R'=phenyl, R1= _-CH2 - _CO2H (i.e. L-aspartic acid), and compounds derived therefrom, wherein R' is either an alkyl group containing up to 4 carbon atoms, or a group of the structure OX, O-oxygen, X=hydrogen, methyl, ethyl, n-propyl, isopropyl or tert-butyl, or A group with the structure -COOY, Y=hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, isobutyl, isopentyl or neopentyl.

In the following approximate but non-limiting examples, preferred R' is the following group:

6,7-Dimethoxynaphthyl (β-Dmn)

5-Methoxynaphthyl (β-Mns)

2,2,5,7,8-Pentamethylchromone (Pmc)

5,6,7,8-tetrahydronaphthyl (Thn)

5,6,7,8-Tetramethylnaphthyl (Tmn)

Phenyl (Phl)

4-Methoxy-2,3,6-trimethylphenyl (Mtr)

2,3,4,5,6-pentamethylphenyl (Pme)

4-methoxy-2,3,5,6-tetramethylphenyl (Mte)

4-Hydroxy-2,3,6-trimethylphenyl (Htr)

4-Carboxyphenyl (Cph)

Particularly effective is the Mtr group, the related structure derived from 4-methoxy-2,3,6-trimethylbenzenesulfonyl chloride.

Suitable C groups are [alpha]-, [beta]-, [gamma]- or [delta]-amino acids and derivatives thereof, and also N-glycosidically linked uronic acids, such as [beta]-D-glucosamine uronic acid.

The compounds of the present invention are also surprisingly characterized by improved tolerability and thus are superior to other compounds from a technical point of view. For example, the LD ₅₀ of the compound β-naphthalenesulfonyl-L-Asp-D, L-Aph-Pip (rat, intravenous injection) is about 120 mg/kg, while the LD ₅₀ of NAPAP is about 20 mg/kg. It has also been demonstrated that the tolerance is further improved when a carboxyl group is added to the R' group of the compounds of the present invention.

Compounds according to the invention having acidic groups have an improved tolerability, manifested in a reduced histamine release and a reduced blood pressure lowering effect.

These compounds, in addition to their strong antithrombin potency, specificity and good tolerance due to improved stability to enzymes such as trypsin and chymotrypsin and to liver and intestinal homogenates, These substances thus become an important class of anticoagulant drugs.

Another advantage of the compounds of the present invention is oral bioavailability, which makes these compounds particularly attractive.

The compounds of the invention are used therapeutically as antithrombotic agents, for preventing thrombosis or for diagnosis.

The present invention also provides derivatives of these substances, especially ester compounds. It is characterized in that the carboxyl group of the amino acid R1 is esterified by fatty alcohols with up to 17 carbon atoms. These esters are prepared from alcohols such as methyl, ethyl, isopropyl, n-propyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, octane, decane, dodeca, hexadecyl, or heptadecanyl alcohols . After derivatization with alcohols, the lipid solubility of the substances according to the invention is improved, which is reflected in a suitable absorption in the intestinal tract.

If there is a hydroxyl group on the R2 or R3 group, it may optionally be esterified with a carboxylic acid such as acetic acid, succinic acid, pivalic acid, hexanoic acid, octanoic acid, decacarbonic acid, dodecanoic acid or decacarbonic acid. Hexacarbonic acid.

The thrombin inhibitor of the present invention may exist in various salt forms due to the basicity of the amidine group. The salt form can have a major effect on the solubility of the compound and its absorption in therapeutic applications. Salts may be formates, acetates, hexanoates, oleates or carboxylates up to 16 carbon atoms, chlorides, bromides, iodides, alkylsulfonates up to 10 carbon atoms dibasic and tribasic carboxylates, such as citrate or tartrate.

Particularly preferred are:

In the compound of formula I, R' is β-naphthyl, R1 is -CH ₂ COOX, X is equal to hydrogen, R2 and R3 together form piperidine.

In the compound of formula I, R' is β-6,7-dimethoxynaphthyl, R1 is -CH ₂ COOX, X is equal to hydrogen, R2 and R3 jointly form piperidine.

In the compound of formula I, R' is β-tetralin, R1 is -CH ₂ COOX, X is equal to hydrogen, and R2 and R3 together form piperidine.

In the compound of formula I, R' is 4-methoxy-2,3,6-trimethylphenyl, R1 is _-CH2COOX , X is equal to hydrogen, and R2 and R3 together form piperidine.

In the compound of formula I, R' is 4-carboxyphenyl, R1 is _-CH2COOX , X is equal to hydrogen, and R2 and R3 together form piperidine.

In the compound of formula I, R' is 4-hydroxy-2,3,6-trimethylphenyl, R1 is _-CH2COOX , X is equal to hydrogen, and R2 and R3 together form piperidine.

In the compound of formula I, R' is 2,2,5,7,8-pentamethylchroman, R1 is _-CH2COOX , X is equal to hydrogen, R2 and R3 together form piperidine.

The compound of formula I is characterized in that X is an alcohol group with a maximum of 17 carbon atoms.

In the compound of formula I, R1 has the structure -(CH ₂ )n-SO ₃ H, n=1-4.

Formula I compound, the structure of R1 is -(CH ₂ )n-PO(OH) ₂ , n=1-4,

The compound of formula I is characterized in that R2 and R3 jointly form 3-hydroxymethylpiperidine.

Compounds of formula I, characterized in that the hydroxyl group is esterified with a carboxylic acid containing up to 17 carbon atoms.

The content of the present invention also relates to the method for preparing the compound of formula I, which is characterized in that the acidic amino acid whose α-position carboxyl group is protected is coupled to the α-amino group of p-cyanophenylalanine amide derivatives, and then the N - α-protecting group, the sulfonyl chloride is coupled to the nitrogen-containing group, the cyano group is converted into an amidino group, and the protecting group present on the third functional group on the acidic amino acid is optionally cleaved off.

For reactions with acidic amino acids, especially amino acids such as aspartic acid (Asp) or glutamic acid (Glu), protected amino acids must be used. It is therefore necessary to cover the N-alpha-functional group with a protecting group. Preferred groups for use as protecting groups are urethane types such as tert-butoxycarbonyl (Boc), benzyloxycarbonyl (Z or Cbo), diphenylisopropoxycarbonyl (Bpoc), dimethoxydimethylbenzyl Oxycarbonyl (Ddz) or 9-fluorenylmethoxycarbonyl (Fmoc). Special attention should be paid to the protection of the carboxyl group on the side chain. The ester groups introduced for this purpose can preferably be cleaved off after the synthesis if desired. If it is desired to cleave off the protecting group of the side chain group after synthesis, it is best to introduce tert-butyl ester. As N-alpha-protecting group there may be mentioned, for example, the basic cleavable Fmoc group or the acid-sensitive Bpoc or Ddz group. If the side chain of the acidic amino function in the final compound should still be esterified, it is preferably used The desired alcohol esterifies the corresponding N-α-Z protected amino acid to the α-tert-butyl ester. The Z group is removed by hydrogenolysis, the corresponding sulfonyl chloride is reacted with the amino acid derivative, the tert-butyl ester is acidically cleaved, and the separated product is coupled to the amino group of the cyanophenylalanine derivative. The sulfonate amine linkage is usually carried out by a standard method, which is to link the corresponding sulfonyl chloride to the amino group of an acidic amino acid. When the R' of the prepared compound contains a phenolic group, it is necessary to carry out corresponding phenolic ether protection, particularly preferably tert-butyl Ether, react with sulfonyl chloride to form a sulfonamide bond, and then use an acidic method to cleavage to obtain the desired final compound.

Free phenolic groups can also be prepared by other known methods. For example with a demethylating agent, particularly preferably boron tribromide in an organic solvent, a preferred solvent being mentioned dichloromethane.

A second possibility is to first make the acidic amino acid into a sulfonamide bond and then condense it with a p-cyanophenylalanine derivative. For the preparation of arylsulfonylamino acid-tert-butyl ester or heteroaromatic ringsulfonylamino acid-tert-butyl ester, in the presence of a base in dimethylformamide, for example in N-methylmorpholine or diisopropylethyl The corresponding sulfonyl chloride is reacted with amino acid-tert-butyl-α-methyl ester in the presence of an amine. The methyl ester is saponified with caustic and attached to the Aph- or cyanophenylalanine derivative via a carbodiimide reaction. The compounds of the present invention having a sulfonic acid group (B=SO ₃ H), preferably having cysteine sulfonic acid, can be prepared from N-alpha-protected cysteine sulfonic acid, at this time the nitrogen of the amino acid is connected to the Boc group, And connect the compound to cyanophenylalanine piperidine. After cleavage of the protecting group, the desired sulfonyl chloride is reacted. The corresponding cysteine compound with a free sulfhydryl group is also conveniently prepared and then reacted with an oxidizing agent such as performic acid to become a sulfonic acid. For this purpose, BocCys (SStBu) or Boc-Cys (Trt) are preferably used. The oxidation reaction proceeds after cleavage of the sulfur protecting group.

Compounds according to the invention which have the radical C as an α-, β-, γ- or δ-amino acid can be prepared according to known methods. The preferred N-protected acidic amino acid (such as aspartic acid), as the protecting group, is preferably introduced into the final sulfonamide structure, coupled with the carboxyl-protected amino acid, so that the α-carboxyl group is to be protected, preferably methyl ester. The coupling reaction is best carried out with a carbodiimide type condensing agent, preferably dicyclohexylcarbodiimide. Hydroxybenzotriazole can be added to the reaction and the solvent is preferably dimethylformamide.

When the separated substance is a methyl ester, the α-position carboxyl group is freed by saponification.

Then these precursors undergo a coupling reaction at the amino group of cyanophenylalanine according to the method already described, and enter the next step of reaction, that is, the amidine group is prepared.

To prepare compounds of the invention wherein group C is an N-glycosidically linked aldonic acid, the N-alpha and C-alpha protected acidic amino acids are first linked to the corresponding protected carbohydrate moieties. This preparation is carried out by the method known in literature, for example A.Klemer etc.; J.Carbohydrate Chemistry, 7 (4), 785-797 (1988) or T.Takeda etc.; Carbohydrate Research, 207, 71-79 (1990), Or R. Walczyna et al; Car-bohydrate Research, 180, 147-151 (1988), or L. Urge et al; Tetrahydron Letters, 32 (9) 3445-3448 (1991). After cleavage of the nitrogen protecting group of the amino acid, the sulfonyl chloride is ligated as described above, and the protecting group of the C-alpha carboxyl group is cleaved off. Then the C-terminal amino group is coupled, and the C-terminus is preferably amidinophenylalanine, especially D-4-amidinophenylalanine-piperidine amide. After deprotection of the aldonic acid, the material is purified by conventional methods, eg, by gel permeation chromatography or ion exchange chromatography.

To prepare the amidino group, the cyanophenylalanine compound was dissolved in pyridine and triethylamine and saturated with hydrogen sulfide. After 2-3 days the solution was stirred with aqueous hydrochloric acid and a precipitate separated. After methylation with a methylating agent (preferably methyl iodide) and reaction with an ammonium salt (preferably ammonium acetate) in alcohol (preferably ammonium formate), a peptide derivative containing an amidine group can be obtained. After acid hydrolysis treatment with trifluoroacetic acid or HCl in acetic acid, the desired final product is obtained. Gel permeation chromatography in methanol on ^R Sephadex LH-20 then gave the pure material. The target compound was confirmed by NMR and mass spectrometry, and its purity was checked by HPLC and thin layer chromatography. For conversion to the desired salt, ion exchange chromatography is preferred. For this purpose, the corresponding compound is bound to a carboxymethylated ion exchange resin (such as CM- ^R Fractogel), eluted with the desired acid, and the final product is crystallized by low-pressure sublimation drying. Other desired salts can be obtained from Acetate is obtained.

To evaluate the activity of the inhibitors of the present invention, tests can be carried out according to different criteria, the preferred method being the determination of Ki values, IC ₅₀ values and in vivo and ex vivo site thromboplastin time (PTT). Specificity assays were determined using _IC50 values for various serine proteases, especially for thrombin _and trypsin. The determination method of the stability of the compound of the present invention is that the test substance is incubated with pure enzyme (preferably trypsin, chymotrypsin or papain) or with liver or intestinal homogenate, and is sampled from the solution at certain time intervals. It can be determined by HPLC. Tests have shown that the compounds claimed in the present invention are stable from a technical standpoint and decompose relatively slowly. As for the claimed compound as a specific and potent thrombin inhibitor with significant antithrombotic ability, it exceeds the low molecular weight inhibitors known so far, and partly has the characteristics of gastrointestinal administration, just like the compound Mtr -L-Asp-D-Aph-Pip as indicated.

The content of the present invention also relates to diagnostic or therapeutic agents containing compounds of formula I.

In addition, the content of the present invention also relates to the method of applying the compound of formula I to the preparation of diagnostic or therapeutic drugs with antithrombotic effect.

Abbreviations:

Aph amidinophenylalanine

NAPAP β-naphthalenesulfonyl-glycyl-D, L-P-amidinophenylalanyl-piperidine

Asp aspartic acid

Asn Asparagine

Glu glutamic acid

Cys(SO ₃ H) cysteine sulfonic acid

β-Dmn 6,7-dimethoxynaphthyl

β-Mns 5-methoxynaphthyl

Pmc 2,2,5,7,8-pentamethylchroman

Thn 5,6,7,8-tetrahydronaphthalene

Tmn 5,6,7,8-tetramethylnaphthyl

Phl Phenyl

Mtr 4-methoxy-2,3,6-trimethylphenyl

Tos 4-Methylphenyl

Pme 2,3,4,5,6-pentamethylphenyl

Mte 4-methoxy-2,3,5,6-tetramethylphenyl

Htr 4-Hydroxy-2,3,6-trimethylphenyl

Cph 4-carboxyphenyl

Z(Cbo) Benzyloxycarbonyl

Boc tert-butoxycarbonyl

Bpoc Diphenylisopropoxycarbonyl

Ddz Dimethoxydimethylbenzyloxycarbonyl

Fmoc fluorenylmethoxycarbonyl

Pip piperidine

OtBu tert-butyl ester

OMe methyl ester

OEt ethyl ester

OiBu Isobutyl Ester (Second Butyl Ester)

OiPr isopropyl ester

OnPe Neopentyl Ester

DC TLC

DCCI Dicyclohexylcarbodiimide

DMF Dimethylformamide

FAB-MS fast atom bombardment mass spectrometry

DIPEA Diisopropylethylamine

HOBt Hydroxybenzotriazole

The following examples further illustrate the present invention

β-naphthalenesulfonyl-L-aspartate-D-p-amidinophenylalanylpiperidine

1. Boc-D-P-cyanophenylalanyl piperidine

50 g (255 mmol) of p-cyanobenzyl bromide, 55 g (255 mmol) of diethyl acetamidomalonate and 2 g of potassium iodide were boiled in 250 ml of anhydrous dioxane; 6 g (260 mmol) of freshly prepared A solution of sodium and ethanol was added dropwise to the above solution. Heating to reflux for another 3 hours, cooling to 80°C, adding 170ml of 3N sodium hydroxide solution within 3 hours, heating at 95°C for 4 hours. After cooling, the pH was adjusted to 1 with 6N HCl, the dioxane was distilled off, and the precipitate was collected by filtration. Add sodium hydroxide to make the pH = 9, extract with 2 times the amount of ethyl acetate, adjust the pH of the aqueous phase to 1 with hydrochloric acid, and crystallize N-acetylcyanophenylalanine, collect the crystals by filtration, wash with water several times, and vacuum dry. The yield was 47g (79.2%),

Purity test: TLC Rf=0.5 (chloroform 50/methanol 10/glacial acetic acid 2.5∥volume ratio.

Dissolve 24 g of the product in 3 N caustic soda in 3 liters of water, adjust the pH to 6-6.5, add 500 mg of acyltransferase to it, and incubate at 37° C. for 4 days. The solution was ultrafiltered to remove the acyltransferase, and the filtrate volume was concentrated to 1 liter. After adjusting the pH to 1, it was extracted several times with ethyl acetate, the organic phase was washed with a small amount of concentrated salt solution, dried over sodium sulfate, and the solvent was evaporated to obtain 8.2 g of N-acetyl-D-cyanophenylalanine (68%). Add 22ml of glacial acetic acid, 4.3ml of concentrated hydrochloric acid and 40ml of water to 8g of the above compound, heat and boil for 24 hours. The hydrolyzed solution was evaporated, and the residual acid was distilled with methanol, and precipitated with methanol-ether, yielding 6.6 g (85%).

5g of D-cyanophenylalanine hydrochloride was dissolved in 14ml of water by adding 7.5ml of diisopropylethylamine. A solution of 6 g of tert-butoxycarbonyloxyimino-2-phenylacetonitrile in 17 ml of dioxane was added thereto, and stirred overnight. 40ml of water and 50ml of ethyl acetate were added. The aqueous phase was separated and the organic phase was extracted several times with 1M potassium bicarbonate. The aqueous phases were combined, washed several times with 10 ml of ether, and then adjusted to pH=3 with hydrochloric acid. Extracted 3 times with ethyl acetate, washed with sodium chloride solution, dried the organic phase with sodium sulfate, and evaporated the solvent to obtain 5.6g (78%) of Boc-D-cyanophenylalanine. 3.26g (10mmol) Boc-D-cyanophenylalanine, 1.49g (11mmol) HOBt and 2.42g (12mmol) DCCI were dissolved in 50ml DMF and stirred for 1 hour. Add 1 ml of piperidine and stir overnight. The precipitated dicyclohexylurea was filtered off, DMF was distilled off, the residue was dissolved in ethyl acetate, and washed three times with potassium bicarbonate, 1M potassium bisulfate and saturated saline solution. After drying over sodium sulfate, the solvent was distilled off to obtain 3.16 g (80%) of Boc-D-cyanophenylalanylpiperidine,

Purity control: TLC Rf = 0.27 (chloroform)

2. D-cyanophenylalanyl piperidine hydrochloride

3g of the Boc-protected compound was dissolved in 50ml of 1.2N HCl in ice-water acetic acid solution, and stirred at room temperature for 30 minutes. The cleavage agent was evaporated in vacuo, evaporated with toluene, and the residue was triturated with a small amount of ether. The crystals were collected by filtration and dried in vacuo. The yield was 2.2 g (90%).

3. Ddz-Asp(tBu)-D-cyanophenylalanyl piperidine

2.88g (7mmol) Ddz-Asp (tBu), 1.04g HOBt, 1.73g DCCI and 2.06g (7mmol) cyanophenylalanyl piperidine hydrochloride were dissolved in 50ml DMF, and 2.4ml (14mmol) diisopropylethyl After amine, stir in the dark at room temperature for 1 day, evaporate the solvent in vacuo, dissolve the residue in ethyl acetate, wash 3 times with 1M potassium bisulfate solution, 3 times with potassium bicarbonate solution and 2 times with concentrated salt solution . After drying the organic phase over sodium sulfate, the solvent is evaporated and the residue is triturated with diisopropyl ether. The crystals were collected by filtration and dried to yield 3.86 g (85%) of Ddz-Asp(tBu)-D-cyano-phenylalanylpiperidine.

4. β-Naphthalenesulfonyl-Asp-(t-Bu)-D-cyanophenylalanylpiperidine

3.25 g (5 mmol) of Ddz-Asp(tBu)-D-cyanophenylalanylpiperidine were dissolved in 200 ml of 5% trifluoroacetic acid in dichloromethane and stirred at room temperature for 30 minutes. The reaction solution was poured into 2M aqueous sodium bicarbonate solution, and the organic layer was washed twice with 2M sodium bicarbonate solution and once with concentrated salt solution. After drying over sodium sulfate, the solvent was evaporated and the oily residue was extracted 3 times with diisopropyl ether. A dichloromethane solution of 1.71 ml (10 mmol) of DIPEA was added to dissolve the oil, and 1.13 g (5 mmol) of β-naphthalenesulfonyl chloride were added thereto with stirring. Stir the reactant at room temperature overnight _and then wash the organic phase with NaHCO solution, potassium bisulfate solution and sodium chloride solution three times respectively, dry the organic phase over sodium sulfate, evaporate the solvent, and obtain the material without purification for the following One step reaction.

5. β-Naphthalenesulfonyl-AsP(tBu)-D-amidino-phenylalanylpiperidine

The compound obtained in 4 was dissolved in 30ml of dry pyridine, 1ml of triethylamine was added, and hydrogen sulfide gas was introduced for 3 hours. After standing at room temperature for 3 days, it was poured into a mixture of 100 g of ice and 50 ml of concentrated hydrochloric acid. The precipitated precipitate was suction filtered and washed with water. After drying, the thioamide was dissolved in 50ml of acetone and mixed with 1.5ml of methyl iodide. Heat to reflux for 30 minutes. After cooling, it was precipitated with ether. The precipitate was dissolved in dichloromethane and washed twice with water. The organic phase was dried over sodium sulfate, the solvent was evaporated, the residue was dissolved in 30 ml of dry methanol, and 200 mg of ammonium acetate was added. After heating at 60°C for 3 hours, the solvent was evaporated in vacuo. The product was purified by chromatography on Sephadex LH-20, eluting with methanol, and the yield was 630 mg.

Purity control: TLC Rf=0.55 (chloroform 50/methanol 10/glacial acetic acid 2.5∥volume).

FAB-MS: M+H 636

6. β-naphthalenesulfonyl-Asp-D-amidino-phenylalanylpiperidine

500 mg of the compound isolated in 5 was dissolved in 5 ml of a solution of 1.2N HCl in glacial acetic acid, and stirred at room temperature for 1 hour. After evaporation of the solvent, the residue was triturated with ether. The solid was collected by filtration and the product was dried over phosphorus pentoxide under high vacuum. The crude product was dissolved in water and bound to CM-fraction gel ion exchange resin. Elution with 5% acetic acid gave 290 mg of acetate after lyophilization. Its correct structure was proved by ¹³ C-NMR spectrum.

MAB-MS: M+H 580

6,7-Dimethoxy-β-naphthalenesulfonyl-L-Asp-D-p-amidinophenylalanylpiperidine

The preparation method is similar to the above example, except that β-naphthalenesulfonyl chloride is replaced by 6,7-dimethoxy-β-naphthalenesulfonyl chloride.

MAB-MS: M+H 640

5,6,7,8-Tetrahydro-β-naphthalenesulfonyl-L-Asp-D-p-amidinophenylalanylpiperidine

With 5,6,7,8-tetrahydro-β-naphthalenesulfonyl chloride and the above method, 5,6,7,8-tetrahydro-β-naphthalenesulfonyl-L-Asp-D-p-amidinophenylpropane Aminopiperidine.

FAB-MS: M+H 584

4-Methoxy-2,3,6-trimethylbenzenesulfonyl-L-Asp-D-P-amidinophenylalanylpiperidine

Using 4-methoxy-2,3,6-trimethylbenzenesulfonyl chloride and the method described above, the preparation of 4-methoxy-2,3,6-trimethylbenzenesulfonyl-L-Asp-D-p- Aminophenylalanylpiperidine.

FAB-MS: M+H 602

4-Methoxy-2,3,6-trimethylbenzenesulfonyl-L-Cys(SO ₃ H)-DP-amidinophenylalanylpiperidine

Using 4-methoxy-2,3,6-trimethylbenzenesulfonyl chloride and the method described above, the preparation of 4-methoxy-2,3,6-trimethylbenzenesulfonyl-L-Cys

(SO ₃ H)-Dp-amidinophenylalanylpiperidine

FAB-MS: M+H 638

4-Methoxy-2,3,6-trimethylbenzenesulfonyl-L-aspartyl-N-[β-D-glucopyranosyl)aldonoic acid]-D-P-amidinophenylalanine Acyl piperidine (Mtr-L-Asp(β-D-aminoglucuronic acid)_D_Aph-Pip)

1. Methyl (2,3,4-tri-O-acetyl-β-D-glucopyranosesamine)aldonate

2 g of (2,3,4-tri-O-acetyl-β-D-glucopyranose azide)-aldonate methyl ester was dissolved in 200 ml of ethyl acetate/methanol (2:1, v/v), Add 2 g of palladium-carbon, adjust the pH to 7.5 with triethylamine, and treat with hydrogen for 1 hour. The reaction mixture was filtered, the solvent was evaporated, and the yield was 1.9g

Purity control: TLC Rf=0.35 (dichloromethane: acetone/2:1)

2. 2-N-(Z)-4-N[(2,3,4-Tri-O-acetyl-β-D-glucopyranosyl)aldonate methyl ester]-L-asparagine-tert butyryl ester

1.26g of Z-aspartic acid-α-tert-butyl ester, 0.9g of HOBt and 1.4g of dicyclohexylcarbodiimide were dissolved in 100ml of THF and mixed with 1.4g of the product of step 1 at 0°C. After stirring overnight at room temperature, the solvent was evaporated in vacuo. The residue was dissolved in chloroform, washed with water, dried over sodium sulfate, and the solvent was distilled off. The residue was purified on silica gel, eluting with chloroform/acetone (6:1/v:v). Yield 1.7g

Purity control: TLC R _f =0.72 (chloroform:acetone/6:1)

3. Methyl 4-N-[(2,3,4-tri-O-acetyl-β-D-glucopyranosyl)aldonate]-L-asparagine tert-butyl ester

0.9 g of the compound obtained in the second step was dissolved in 20 ml of methanol, mixed with a small amount of palladium-carbon, hydrogenated for 5 hours, the catalyst was filtered off, the solvent was evaporated, and the residue (0.75 g) could be used in the next step without further purification.

4.2-N-(4-methoxy-2,3,6-trimethylbenzenesulfonyl)-4-N-[(2,3,4-tri-O-acetyl-β-D-pyran Glucosyl)-aldonate methyl ester]-L-asparagine-tert-butyl ester

1.73g of the above substance, 1.2ml of diisopropylethylamine and 0.8g of Mtr-chloride were dissolved in 80ml of DMF, stirred overnight at room temperature, the solvent was evaporated in vacuo, the residue was dissolved in ethyl acetate, washed three times with water, dried over sodium sulfate After phase, the solvent was evaporated in vacuo. It was then purified by silica gel column chromatography, eluting with dichloromethane-acetone (3:1/v/v). The yield was 1.57g.

Purity control: TLC Rf=0.87 (dichloromethane:methanol/10:1)

5.2-N-(4-methoxy-2,3,6-trimethylbenzenesulfonyl)-4-N-[(2,3,4-tri-O-acetyl-β-D-pyran Glucosyl)aldonate methyl ester]-L-asparagine

2.2 g of the substance from step 4 was dissolved in 50 ml of a mixture of trifluoroacetic acid/dichloromethane (1:1/v:v) and stirred at room temperature for 1 hour. The acid mixture was evaporated in vacuo, followed by toluene to remove traces of the acid present, and the product obtained was used in the next reaction without further purification. The yield was 1.2g.

Purity control: TLC Rf=0.7 (chloroform:methanol/3:1)

6.2-N-(4-methoxy-2,3,6-trimethylbenzenesulfonyl)-4-N-[(2,3,4-tri-O-acetyl-β-D-pyran Glucosyl)aldonate methyl ester]-L-aspartyl-D-4-amidinophenylalanylpiperidine

1.0g of the above substance, 0.23g of HOBt and 0.37g of DCCI were dissolved in 50ml of DMF, stirred at 4°C for 30 minutes, then added 0.4g of D-4-amidinophenylalanylpiperidine and 0.5g of N-methyl phylloline. The reaction was stirred overnight at room temperature. After filtration, the solvent was evaporated in vacuo. The crude product was chromatographed on silica gel, eluting with chloroform/methanol 5:1, and the yield was 1.2 g.

Purity control: TLC Rf=0.7 (chloroform:methanol/3:1)

7. 2-N-(4-Methoxy-2,3,6-trimethylbenzenesulfonyl)-4-N-[(β-D-glucopyranosyl)aldonate methyl ester]-L-day Partyyl-D-4-amidinophenylalanylpiperidine

1.1 g of the substance from the previous step was dissolved in 50 ml of methanol, adjusted to pH = 9.2 hours with 1N sodium hydroxide, neutralized with methanolic HCl at pH = 8.5-9, and the solvent was evaporated in vacuo. The residue was purified by chromatography on ^R Sephadex LH-20 in methanol. The yield was 0.9g.

Purity control: TLC Rf=0.34 (chloroform:methanol:water/40:20:1).

8. 2-N-(4-methoxy-2,3,6-trimethylbenzenesulfonyl)-4-N-(β-D-glucopyranosyl)aldonoic acid-L-aspartyl- D-Amidinophenylalanylpiperidine

0.9g of the substance from the previous step was dissolved in 100ml of chloroform:methanol:water (40:20:1/v:v:v), mixed with 0.2g of barium hydroxide, the mixture was stirred at room temperature for 3 hours, and adjusted to pH with methanolic HCl = 3.5, and the solvent was distilled off. The residue was purified by chromatography on Sephades ^R LH-20 in methanol. The yield was 0.72g.

Purity control: TLC Rf=0.32 (chloroform:methanol:water/8:6:1)

FAB-MS: 777

4-Methoxy-2,3,6-trimethylbenzenesulfonyl-L-aspartic acid-(γ-aminobutyric acid)-D-Aph-piperidine

1.4-Methoxy-2,3,6-trimethylbenzenesulfonyl-L-aspartic acid-(tert-butyl γ-aminobutyrate)-D-cyanophenylalanylpiperidine

1gMtr-Asp-D-cyanophenylalanylpiperidine was dissolved in 30ml DMF, and then mixed with 0.39ml diisopropylethylamine, 0.39gHOBt, 0.42gγ-aminobutyric acid tert-butyl ester and 0.66g dicyclohexyl carbon di imine mix. Stir in the ice bath for 1 hour and then at room temperature overnight. The precipitated dicyclohexylurea was filtered off and the solvent was evaporated in vacuo. The oily residue was dissolved in ethyl acetate and washed twice with water, 1M potassium bicarbonate solution, 1M potassium bisulfate solution and saturated sodium chloride solution respectively. After drying over sodium sulfate, the solvent was evaporated, yield: 1.1 g.

Purity control: TLC Rf = 0.89 (chloroform:methanol/9:1)

2.4-Methoxy-2,3,6-trimethylbenzenesulfonyl-L-aspartic acid-(tert-butyl γ-aminobutyrate)-D-Aph-piperidine

According to the fifth step of Example 1, the cyano group was converted into an amidino group, and the yield: 500 mg.

Purity control: TLC Rf=0.4 (chloroform:methanol:acetic acid/50:10:2.5)

3.4-Methoxy-2,3,6-trimethylbenzenesulfonyl-L-aspartic acid-(γ-aminobutyric acid)_D-Aph-piperidine

400 mg of the above-prepared substance was dissolved in 15 ml of 1.2N HCl/acetic acid and stirred at room temperature for 90 minutes. The acid mixture was distilled off, mixed twice with toluene and distilled off. The crude product was dissolved in 3 ml of methanol and dropped into 50 ml of ether to crystallize it. The precipitate was collected and dried in vacuo. Yield: 320 mg.

Purity control: TLC Rf=0.25 (chloroform:methanol:acetic acid/50:10:2.5).

FAB-MS: 686

Racemate resolution using acyltransferase in step 1 converts the corresponding racemic D,L-cyanophenylalanylpiperidines to compounds of the invention, which are also very potent thrombin inhibitors.

Table 1 General list of compounds of the present invention

β-Naphthalenesulfonyl-L-Glu-D-Aph-Pip

-L-Asp-D-Aph-Pip

-L-Asp-D-Aph-Pip

-D-Asp-D，L-Aph-Pip

-D-Asp-D, L-Aph-Pip

-L-Asp-D，L-Aph-Pip

-L-Asp-D, L-Aph-Pip

-L-Asp-D，L-Aph-3-羟甲基哌啶

-L-Asp-D, L-Aph-3-hydroxymethylpiperidine

-L-Asp（OtBu）-D-Aph-Pip

-L-Asp(OtBu)-D-Aph-Pip

-L-Asp(OMe)-D-Aph-Pip

-L-Asp（OEt）-D-Aph-Pip

-L-Asp(OEt)-D-Aph-Pip

-L-Asp（OiBu）-D-Aph-Pip

-L-Asp(OiBu)-D-Aph-Pip

Mtr-D-Asp-D, L-Aph-Pip

Mtr-L-Asp(OtBu)-D-Aph-Pip

Mtr-L-Asp(OMe)-D-Aph-Pip

Mtr-L-Asp(OEt)-D-Aph-Pip

Mtr-L-Asp(OiBu)-D-Aph-Pip

Mtr-L-Asp(OiPr)-D-Aph-Pip

Mtr-L-Asp-D-Aph-Pip

Mte-L-Asp-D-Aph-Pip

Mte-L-Asp(OiBu)-D, L-Aph-Pip

Htr-L-Asp(OtBu)-D-Aph-Pip

Htr-L-Asp-D-Aph-Pip

Mtr-L-Glu-D, L-Aph-Pip

Mtr-L-Asn-D-Aph-Pip

Mtr-D-Asn-D, L-Aph-Pip

Thn-L-Asn-D-Aph-Pip

Pme-L-Asp(OtBu)-D-Aph-Pip

Pme-L-Asp-D-Aph-Pip

Pme-L-Asp(OtBu)-D, L-Aph-Pip

Phl-L-Asp(OtBu)-D-Aph-Pip

Phl-L-Asp-D-Aph-Pip

B-Dmn-L-Asp(OtBu)-D-Aph-Pip

Table 1 (continued)

B-Dmn-L-Asp-D-Aph-Pip

Tos-L-Asp(OtBu)-D-Aph-Pip

Tos-L-Asp-D-Aph-Pip

Pmc-L-Asp(OtBu)-D-Aph-Pip

Pmc-L-Asp-D-Aph-Pip

B-Mns-L-Asp(OnPe)-D-Aph-Pip

B-Mns-L-Asp-D-Aph-Pip

Cph-L-Asp(OtBu)-D, L-Aph-Pip

Cph-L-Asp-D, L-Aph-Pip

Phl-L-Cys( _SO3H )-D-Aph-Pip

Mtr-L-Cys( _SO3H )-D, L-Aph-Pip

Mtr-L-Cys( _SO3H )-D-Aph-Pip

Mtr-L-Asp (r-aminobutyric acid)-D-Aph-Pip

Mtr-L-Asp (L-Threonine)-D-Aph-Pip

Mtr-L-Asp (L-Phenylalanine)-Aph-Pip

Mtr-L-Asn[(B-D-glucopyranosyl)-aldonic acid]-D-Aph-Pip

* The amidinate salt is not listed.

Determination of thrombin inhibition constant

Inhibition constants (Ki) of substances are determined by known kinetic methods of enzymology. The human thrombin used was 87% pure by means of "active site titration". Ki-assay is composed of buffer (50mM Tris-Hcl, 75mMNaCl pH 7.8, 37°C), 100pM thrombin, 0.1mM substrate S2238 (Kabi) and inhibitor, the concentration range of inhibitor is between 0-0.2μM between. The inhibitor was pre-incubated with the enzyme for 10 minutes and the reaction was initiated by the addition of the chromogenic substrate S2238. To estimate the kinetics, the Ki values (Table II) and the type of inhibition were determined by means of non-linear regression using a "close-knit" mathematical algorithm. The inhibition pattern for all inhibitors proved to be greater than 90% competitive inhibition.

Determination of Inhibitor Specificity

Inhibitor specificity was determined against thrombin and trypsin. Specificity was defined by the ratio of Ki values for trypsin and thrombin (Table II). The inhibitor concentration at which 50% inhibition of enzyme activity occurs is defined as the IC ₅₀ (100% corresponds to an uninhibited enzyme reaction). _IC50 values for trypsin were calculated by preincubating trypsin from bovine pancreas with increasing concentrations of inhibitor in 25 mM Tris-Hcl, 10 mM _CaCl2 , pH=7.8 at 37°C for 10 minutes. The substrate Chromozym TRY (7.1 x 10 ^-5 M) was added to initiate the enzyme reaction. Released pNA was measured at 405 nm after 1 hour. The IC ₅₀ for thrombin was determined by the same method, except that human thrombin was used, the buffer was 50 mM Tris-Hcl, 50 mM Nacl, pH=7.8, and 5×10 ^-5 M substrate S2238. Ki values can be calculated from the IC ₅₀ of thrombin and trypsin (Ki = IC ₅₀ /S + Km). The Ki ratio of trypsin to thrombin is the specificity, and the results are listed in Table II.

Table 2 Effects of some selected compounds compound specific thrombin/trypsin Thrombin inhibition K I (nanomol/L) 1.Napap2.Nas-Asp-D-Aph-pip3.Dmn-Asp-D-Aph-pip4.Mtr-Asp-D-Aph-pip5.Mtr-Asp(tBu)-Aph-pip6.Mtr-Asp(iPr )-Aph-pip7.Nas-Asn-Aph-pip8.Mtr-Asn[(β-glucopyranosyl)aldonic acid]D-Aph-pip9.Mtr-L-Cys(so ₃ H)-D-Aph -pip10.Mtr-l-Asp (r-aminobutyric acid) D-Aph-pip 2472301252973784841061501810 111061.50.90.660.0850.60.8

Claims (14)

1, the method for preparing formula I compound

in R' is a naphthalene ring connected to the α-position or the β-position, and the ring may be optionally substituted by an alkyl group having up to 3 carbon atoms and/or an alkoxy group containing up to 3 carbon atoms; or A tetralin ring or an indene ring attached to the α- or β-position, optionally containing an alkyl group with up to 3 carbon atoms, and/or an alkoxy group with up to 3 carbon atoms; or It is a benzene ring, optionally containing an alkyl group with up to 4 carbon atoms on the ring, and/or having up to three groups with the structure O-X, where O is oxygen, X is hydrogen, methyl, ethyl, normal Propyl, isopropyl or tert-butyl, and/or groups having the structure -COOY, Y is hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, isobutyl, iso Pentyl or neopentyl; or a chroman ring with up to 5 alkyl groups and each alkyl group can have up to 3 carbon atoms; R1 is a group with the structure AB, A=-(CH ₂ )n-, n=1-4, B is an acid group, which can be a carboxylic acid group, which can be optionally esterified or become an amide, and ester alcohols are the most It can contain 17 carbon atoms; it can be a sulfonic acid group, a phosphoric acid group, a boronic acid group or a tetrazolyl group; R1 or a group with the structure ABC, A has the same meaning as above, B is a carbonyl or sulfonyl group, and C is an N-linkage α, β, γ or δ-amino acids or aldonic acid groups linked via N-glycosidic bonds; R2 and R3 may be the same or different, and are alkyl groups with up to 4 carbon atoms, or R2 and R3 together form a heterocyclic ring with up to 8-membered rings, and the rings may be substituted by hydroxyl groups or have up to 3 carbon atoms Substituted by hydroxyalkyl group, the hydroxyl group can be optionally esterified, the corresponding carboxylic acid can contain up to 17 carbon atoms, * represents the carbon atom in the R or S configuration, the R configuration is preferred, The preparation method is characterized in that the protected acidic amino acid is connected to the α-amino group of the p-cyanophenylalanine derivative through the carboxyl group at the α-position, the -α-protecting group is removed, and the sulfonyl chloride is connected to the nitrogen-containing group On the cyano group, the cyano group is converted to an amidino group, and optionally the protecting group present on the third functional group on the acidic amino acid is cleaved off. 2. A process as claimed in claim 1, characterized in that compounds in which * is a carbon atom in the R configuration are prepared. 3. A method according to claim 1, characterized in that the compound wherein R' is β-naphthyl, R1 is -CH ₂ COOX where X is hydrogen, and R2 and R3 jointly form piperidine is prepared. 4. The method of claim 1, characterized in that the preparation of R' is β-6,7-dimethoxynaphthyl, R1 is _-CH2COOX (X=H), R2 and R3 jointly form a compound of piperidine . 5. A process according to claim 1, characterized in that R' is β-tetralin, R1 is _-CH2COOX (X=H), and R2 and R3 together form piperidine. 6. The method of claim 1, characterized in that the preparation of R' is 4-methoxy-2,3,6-trimethylphenyl, R1 is -CH ₂ COOX (X=H), R2 and R3 are collectively Compounds that form piperidines. 7. A process according to claim 1, characterized in that a compound is prepared wherein R' is 4-carboxyphenyl, R1 is _-CH2COOX (X=H), and R2 and R3 together form piperidine. 8. The method of claim 1, characterized in that the preparation of R' is 4-hydroxy-2,3,6-trimethylphenyl, R1 is -CH ₂ COOX (X=H), and R2 and R3 together form a piperidine ring compound of. 9. The process of claim 1, characterized in that R' is prepared as 2,2,5,7,8-pentamethylchroman, R1 is _-CH2COOX (X=H) and R2 is combined with R3 together form the compound of piperidine. 10. The method according to any one of claims 1-8, characterized in that X is an alcohol group, and the alcohol group can have up to 17 carbons. 11. Process according to any one of claims 1-8, characterized in that compounds wherein R1 is -(CH ₂ )n-SO ₃ H (n=1-4) are prepared. 12. Process according to any one of claims 1-8, characterized in that compounds wherein R1 is -(CH ₂ )n-PO(OH) ₂ (n=1-4) are prepared. 13. Process according to any one of claims 1-10, characterized in that compounds in which R2 and R3 together form 3-hydroxymethylpiperidine are prepared. 14. Process according to claim 13, characterized in that compounds are prepared in which the hydroxyl group is esterified with a carboxylic acid containing up to 17 carbon atoms.